b r a z i l i a n j o u r n a l o f m i c r o b i o l o g y 4 9 (2 0 1 8) 104–111

http://www.bjmicrobiol.com.br/

Food Microbiology

Growth kinetics of Escherichia coli O157:H7 on the

epicarp of fresh vegetables and fruits

Mariel Gullian-Klanian ∗ , Maria José Sánchez-Solis

University Marist of Mérida, Experimental Research Unit, Yucatán, Mexico

a r t i c l e i n f o a b s t r a c t

Article history:
Received 17 September 2015
Accepted 20 October 2016
Available online 30 August 2017
Associate Editor: Beatriz Ernestina
Cabilio Guth
Despite the increasing reports on the incidence of fresh vegetables and fruits as a possible
vehicle for human pathogens, there is currently limited knowledge on the growth potential
of Escherichia coli O157:H7 on different plant substrates. This study analyzed the selective
adhesion and growth of E. coli O157:H7 on chili habanero (Capsicum chinense L.), cucumber
(Cucumis sativus), radish (Raphanus sativus), tomato (Lycopersicon esculentum), beet (Beta vul-
garis subsp. vulgaris), and onion (Allium cepa L.) under laboratory conditions. The Gompertz
parameters were used to determine the growth kinetics. Scanning electron microscopy was

Keywords: used to visualize the adhesion of E. coli O157:H7 on the epicarp of the samples. Predictive

E. coli O157:H7 models were constructed to compare the growth of E. coli O157:H7 on the samples with dif-
Adhesion ferent intrinsic factors and to demonstrate the low selectivity of the pathogen. No significant
Predictive growth difference was observed in the lag-phase duration (LPD), generation time (GT), and expo-
Low selectivity nential growth rate (EGR) of the pathogen adhered to the samples. The interaction between
Fruit and vegetables the microorganism and the substrate was less supportive to the growth of E. coli O157:H7
for onion, whereas for tomato and cucumber, the time for the microorganism to attain the
maximum growth rate (M) was significantly longer than that recorded for other samples.
© 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. This is
an open access article under the CC BY-NC-ND license (http://creativecommons.org/
licenses/by-nc-nd/4.0/).

from animal reservoirs (cattle, goats, and sheep) to humans

Introduction through a variety of food vehicles that plays a critical role
in its pathogenicity. E. coli O157:H7 employs multiple mech-
Escherichia coli O157:H7 is a foodborne zoonotic pathogen that
anisms for colonizing vegetables through adhesion to cell
serving as the causative agent of hemorrhagic colitis and
surfaces. The microorganism adheres strongly to tomato skin,
hemolytic uremic syndrome (HUS) in humans. This microor-
spinach leaves and alfalfa sprout roots by curli,1,2 which is
ganism possesses a remarkable capability to get transmitted
mediated by the filamentous type III secretion system com-
posed of EspA filaments.3 Flagella also play a key role in the
leaf attachment process in E. coli O157:H7.4 The internaliza-
tion of E. coli O157:H7 has been demonstrated under controlled
∗
Corresponding author. environmental conditions in the seedlings of temperate
E-mail: mgullian@marista.edu.mx (M. Gullian-Klanian).
https://doi.org/10.1016/j.bjm.2017.08.001
1517-8382/© 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. This is an open access article under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
 b r a z i l i a n j o u r n a l o f m i c r o b i o l o g y 4 9 (2 0 1 8) 104–111
vegetable crops such as lettuce, tomato, cress, radish, alfalfa,
and mung bean,5–8 and the adhesion and internalization of
E. coli O157:H7 in mature leafy vegetables have been demon-
strated both experimentally and in field conditions.9–13
Despite the increasing incidence of fresh produce as a
vehicle for human pathogens, there is a dearth of informa-
tion on the growth potential of E. coli O157:H7 on different
plant substrates. Although most of the outbreaks involving the
occurrence of E. coli O157:H7 on leafy vegetables have been
associated with cross-contamination14–17 there are very few
studies that have investigated the adhesion and growth of
E. coli O157:H7 in other mature vegetables.18–22 Interestingly,
many of these vegetables possess a variety of intrinsic fac-
tors that can considerably reduce the growth of foodborne
microorganisms; however, E. coli O157:H7 manages to success-
fully survive under these varying conditions.
Assuming that the degree of mutual interaction between
bacterial strain and the specific type of plant affects their
association, this study aimed to investigate the capacity of
E. coli O157:H7 to colonize freshly consumed vegetables and
fruits under experimental conditions. The growth of the
pathogen on different substrates was compared using the
Gompertz equation to obtain bacterial growth curves and
linear regression-derived parameters such as the lag-phase
duration, maximum microbial population, and specific micro-
bial growth.
Materials and methods
Microorganism and preparation of inoculums
Escherichia coli O157:H7 strain (SS11) used in this study
was isolated from the feces of pigs from Santa Elena
in Yucatán, Mexico.23 The strain was isolated after pre-
enrichment with EC broth (Difco, Detroit, MI, USA) using
the immunomagnetic separation procedure (IMS) employ-
ing anti-E. coli O157 Dynabeads (Dynal Biotech, Norway)
as described by the manufacturer. The IMS-enriched bacte-
rial culture (50 ␮L) was streaked onto Chromocult coliform
agar (Merck, Darmstadt, Germany) and Sorbitol MacConkey
agar (CM813; Oxoid Ltd, USA) supplemented with cefixime-
potassium tellurite (SR172; Oxoid Ltd.). The phenotype of the
microorganism was characterized with the API 20E system
(BioMérieux, France). Strain serotype O157:H7 was identi-
fied with the immunocapture system and latex agglutination
(Tecra Diagnostics, NSW, Australia). Polymerase chain reac-
tion (PCR) was used to confirm the presence of rfbO157 and
stx1 genes in the strain SS11 and to confirm the absence
of stx2 verotoxin gene. For this purpose, three pairs of
oligonucleotide primers were synthesized for the amplifica-
tion of rfbO157 (420 bp), stx2 (584 bp) and stx1 (306 bp).24–26
The PCR products were detected by 2% agarose gel-
electrophoresis at 100 V and staining with 10 ␮g/mL ethidium
bromide.
The suspensions of E. coli (104 CFU/mL) were prepared for
the inoculation of the vegetables and fruits. A single colony
of E. coli O157:H7 from a streak plate was suspended in
20 mL of EC selective enrichment broth (Difco) and incubated
overnight at 37 ◦ C (16–18 h) in a shaking incubator at 150 rpm
105
(Thermo Scientific, USA). The overnight culture (1 mL) was
inoculated into 99 mL of EC broth and incubated at 37 ◦ C till
it attained the exponential phase (105 CFU/mL). The bacterial
cell concentrations were estimated by comparing the trans-
mittance of the cell suspension with a previously determined
standard curve at 540 nm using a UV–visible spectrophotome-
ter. The final concentration of 104 CFU/mL was obtained by
diluting the bacterial suspension with 400 mL of EC broth,
and the concentration of the inoculum was further con-
firmed by plating 0.1 mL of an appropriately diluted culture
on TSA.
Selection and preparation of vegetables and fruits
The prepared E. coli O157:H7 suspension was inoculated in
triplicate on three vegetables viz., radish (Raphanus sativus),
beet (Beta vulgaris subsp. vulgaris), and onion (Allium cepa L.),
and on three fruits viz. ripe red tomato (Lycopersicon esculen-
tum L.), cucumber (Cucumis sativus), and green habanero chili
(Capsicum chinense L.). Fresh and healthy-looking fruit and veg-
etable samples were purchased no more than one day prior
to the inoculation from a local supermarket and stored at
10 ◦ C till use. The samples with visible defects on the surface
such as bruises, cuts, or abrasions were excluded from the
experiment. The vegetables were decontaminated by immers-
ing in 200 mg/L of chlorine solution (6.5 mg/L of free chlorine)
for 2 min before inoculation. The decontamination procedures
were previously standardized based on the absence of growth
of Enterobacteriaceae on MacConkey agar. The samples were
then rinsed with sterile deionized water for 1 min and air dried
®
with paper towels in a biosafety cabinet (Esco, Infinity Class
II, Australia).
Growth of E. coli O157:H7 on the epicarp layer of fruits
and vegetables
The samples were treated for five different periods (0, 3, 5,
7 and 9 h) of incubation to determine the adhesion capacity
of the E. coli O157:H7. The samples were completely sub-
merged into 400 mL of E. coli O157:H7 broth (104 CFU/mL) for
30 min and gently shaken at 100 rpm at 37 ◦ C. Then the sam-
ples were kept individually in sterile air bubble polyethylene
®
bags (Stomacher Bag, Seward, UK) for the defined period of
time for each treatment. After incubation, the samples were
washed thrice with sterile deionized water and air dried in a
biosafety cabinet. In addition, two control samples were con-
sidered. The first one was an internal control of inoculated
samples that were not subjected to the final washes, and sec-
ond one was a negative control of inoculated samples that
were decontaminated with 200 mg/L of chlorine solution in
the final wash.
The adhesion of E. coli O157:H7 to the epicarp of vegeta-
bles and fruits was determined by spreading the samples on
MacConkey agar. Three slices of the vegetable shell (10 g) were
cut off aseptically and soaked in 90 mL of phosphate buffered
saline (PBS, pH 7.1). After a five-fold serial dilution, 0.1 mL of
the sample solution was inoculated on MacConkey agar in
duplicate and incubated for 24 h at 35 ± 1 ◦ C. The number of
colonies observed in the plates was used to calculate the CFU/g
of tissue.
106 b r a z i l i a n j o u r n a l o f m i c r o b i o l o g y 4 9 (2 0 1 8) 104–111
Proximate analysis
AOAC (Association of Analytical Communities) official
methods27 were used for analysis of moisture (Method
925.09), total protein (Method 950.48), fat (Method 983.23),
and ash contents of the samples (Method 930.05). pH val-
ues were measured by placing the flat electrode (Sensorex,
Stanton, USA) directly against the surface of the exposed
tissue at three different points. Proximal analysis of the
epicarp was performed using the skin of the fruits and
vegetables.
Scanning electron microscopy (SEM)
The adhesion of E. coli O157:H7 to the exocarp of the sam-
ples was visualized using a scanning electron microscope
(Philips model XL 30 SEM microscope) operating at an accel-
erating voltage of 15 kV. A final pressure of 4 Torr (533.3 Pa)
was gradually generated in the chamber in order to pre-
vent the samples from drying, prior to performing the
analysis.28
Modeling of microbial growth
All the experiments were performed in triplicate. The micro-
bial counts per gram of the sample were converted to log10
values before the calculations. The mathematical modeling
of the microbial growth (predicted data) was performed by
employing the modified Gompertz equation:
L(t) = A + C e−e(−B(t−M)) ,
where L(t) is the log of the number of bacteria at time
‘t’ (in hour). A is the asymptotic log of number of bacte-
ria as ‘t’ decreases indefinitely; C is the asymptotic amount
of growth (log number) that occurs as ‘t’ increases indef-
initely; M is the time (in hour) at which the absolute
growth rate is at maximum; and B is the relative growth at
time ‘M’.29,30
The equation was fitted using a nonlinear regression
routine of the SYSTAT statistical computer system package
(SYSTAT Inc., Evanston, IL, USA). The algorithm was selected
from the set of parameters with the lowest residual sum
of squares with a 95% confidence interval. The Gompertz
parameters (A, B, C, and M) were subsequently used to
calculate the growth kinetics, including the exponential
growth rate, EGR (log CFU/g/h) = BC/e; the lag-phase dura-
tion, LPD (h) = M − 1/B; the generation time, GT (h) = log(2)e/BC;
and the maximum population density, MPD (log CFU/g)
= A + C.
Statistical analysis
An analysis of variance (ANOVA) was performed to compare
the growth parameters of E. coli O157:H7 obtained from the
growth curves. The ANOVA and least significance difference
(LSD) between the means were calculated at 95% confidence
level using the SYSTAT software. The means that differed from
the calculated LSD were considered statistically significant
(˛ = 0.05).
Results and discussion
This study investigated the quantification of the growth of
E. coli O157:H7 on vegetables and fruits. Although the major
disease outbreaks attributed to this pathogen have been asso-
ciated with leafy vegetables, the results of this study showed
that E. coli O157:H7 is also capable of adhering to vegetables
with different physicochemical characteristics (Fig. 1). The
methodology used in this study for the adhesion of E. coli
O157:H7 was validated through internal controls. The bacte-
rial cell that adhered to the surface of the samples formed
a matrix with the epidermal layer (biofilm), which provided
them adequate support to counter the effects of washing.
E. coli O157:H7 strain possesses several fimbrial and non-
fimbrial adhesins which usually do not play any role in its
pathogenesis, but are involved with its adhesion to surfaces.31
For example, curli-expressing thin-aggregative fimbriae are
responsible for binding to eukaryotic extracellular matrix pro-
teins and promoting the formation of E. coli O157:H7 biofilms
on inert surfaces.32,33 According to Torres et al.,31 the forma-
tion of biofilms increases the probability of survival of E. coli
O157:H7, and hence, facilitates its protection against adverse
environmental conditions. The samples from the vegetables
and fruits that were not subjected to the final wash prior to
plating (internal control) recorded higher bacterial concentra-
tion than those that were washed, suggesting the presence
of both adhered and free microorganism on the surface of
the samples. The SEM images of the samples obtained after
the final washing clearly showed a layer of capsule bound to
the cell wall of the samples that could possibly be associated
with the exopolysaccharides secreted by the microorganism
to form the matrix structure (Fig. 1D and E).
The Gompertz function fits well for the observed data with
the sum of squares being less than 1 in all the cases (Fig. 2).
Despite of the different intrinsic factors of the vegetables
and fruits used in this study that could influence the growth
of human pathogenic microorganisms, no significant differ-
ence was found in the lag-phase duration (LPD = 2.17 ± 1.03 h;
mean ± S.E.), generation time (GT = 0.47 ± 0.07 h), or exponen-
tial growth rate (EGR = 0.72 ± 0.12 log CFU mL/h) of the E. coli
O157:H7 adhered to the samples (Table 1). Additional informa-
tion on the proximate composition of the vegetables and fruits
under investigation are shown in Table 2. These values are
comparable to those reported by Kim et al.,34 for perilla leaves
inoculated with E. coli O157:H7 at 36 ◦ C (LPD = 2.40 ± 0.32 h;
EGR = 0.55 ± 0.11 CFU/mL/h). The growth of the E. coli O157:H7
strain on the epicarp of the vegetables and fruits was slower
than that observed under controlled laboratory conditions.
The growth model of Buchanan and Klawitter,35 which con-
sidered the same temperature and pH conditions employed
in this study (35 ± 1 ◦ C; pH 6.0) displayed faster kinetic val-
ues than those observed in this study (LPD = 1.54 h; GT = 0.33 h;
and EGR = 0.92 log CFU/mL/h). This could be an outcome of the
variation in the interactions that occur between the substrate
and pathogen which might not necessarily occur in a growth
medium. Similarly, the duration of the lag phase is dependent
on a wide variety of factors, including the initial concentration
of the inoculum, the time required to recover from the physical
damage or shock due to the transfer, and the time required for
 b r a z i l i a n j o u r n a l o f m i c r o b i o l o g y 4 9 (2 0 1 8) 104–111 107

A B

C D

E
F

Fig. 1 – Scanning electron micrographs demonstrating the adhesion of Escherichia coli O157:H7 on the epicarp of vegetables
and fruits, (A) beet, Beta vulgaris subsp. vulgaris (1550×); (B) habanero Chili, Capsicum chinense L., (1810×); (C) tomato,
Lycopersicon esculentum (1500×); (D) radish, Raphanus sativus (4499×); (E) cucumber, Cucumis sativus (3200×); (F) onion, Allium
cepa L. (1635×).

the synthesis of essential coenzymes or division factors and
other enzymes involved in the metabolism of the substrates
present in the medium.36 In general when pathogens colo-
nize plant surfaces, they are required to evade various defense
mechanisms of the host plants. Plants possess a variety of
complex immune responses, which enable them to detect and
initiate counter measures to control undesired proliferation of
foreign substances.37 In addition, ethylene, a plant hormone,
plays a vital role in the defense against plant pathogens.38
Many microorganisms considerably increase the expression of
ethylene; however the transcription level of ethylene induced
by E. coli O157 were ten-fold lower, suggesting that either the
plants do not sense the presence of the microorganism or
that the microorganism evade the defense mechanisms of the
plants consequently inhibiting the activation of ethylene.39
E. coli O157:H7 growth was lower in onion than other
samples suggesting an interaction that is not completely
favorable between the microorganism and the epicarp of the
vegetable. In the case of tomato and cucumber, the time for
the microorganism to reach the maximum growth rate (M)
was significantly longer than that observed for other samples;
the maximum population densities (MPD) for these vegetables
were also the lowest (Table 1). The variation in the size of the
bacterial populations among the different species has been
attributed to a range of plant characteristics, including the
leaf water content, pH, and the presence of bacterial inhibitory
compounds. In addition, it occurs only when the water pres-
sure on the surface of the product overcomes the internal gas
pressure, and also it depends on the hydrophobic nature of
the surface.40–42 E. coli grows in a broad pH range of 4.4–10.0
with an optimum pH of 6–7.43 Specifically, E. coli O157:H7 can
tolerate harsh acidic conditions, and few other strains have
been reported to be able to survive even at pH 2.5–3.0 for over
4 h.44 Arnold and Kaspar,45 found that E. coli O157:H7 becomes
108 b r a z i l i a n j o u r n a l o f m i c r o b i o l o g y 4 9 (2 0 1 8) 104–111

7.00 7.00

E. coli O157:H7 (Log CFU/mL)

E. coli O157:H7 (Log CFU/mL)
6.50 A 6.50 B
6.00 6.00
5.50 5.50
5.00 5.00
4.50 4.50
4.00 4.00
3.50 3.50
3.00 3.00
2.50 2.50
2.00 2.00
0 3 5 7 9 0 3 5 7 9
Hours Hours
E. coli O157:H7 (Log CFU/mL)

7.00 7.00

E. coli O157:H7 (Log CFU/mL)

6.50 C 6.50 D
6.00 6.00
5.50 5.50
5.00
5.00
4.50
4.50
4.00
4.00
3.50
3.50
3.00
2.50 3.00
2.00 2.50
0 3 5 7 9 2.00
0 3 5 7 9
Hours
Hours

7.00 7.00
E. coli O157:H7 (Log CFU/mL)

E. coli O157:H7 (Log CFU/mL)

6.50 E 6.50 F
6.00 6.00
5.50 5.50
5.00 5.00
4.50 4.50
4.00 4.00
3.50 3.50
3.00 3.00
2.50 2.50
2.00 2.00
0 3 5 7 9 0 3 5 7 9
Hours Hours

Fig. 2 – Observed and predicted data for Escherichia coli O157:H7 adhered to the epicarp of vegetables and fruits, (A) tomato;
(B) radish; (C) cucumber; (D) beet; (E) habanero chili; (F) onion (: internal control; : predicted data; : observed data).

more tolerant to acid when it is in the stationary growth phase
or is starved during the log-phase of growth. The systemic pH
of the vegetables and fruits used in this study is in a suitable
range suitable for the growth of the pathogenic microorgan-
ism (Table 2), tomato and onion possess the lowest pH (4.77
and 5.40, respectively). This suggests that the pH in tomatoes
might have influenced the growth rate of E. coli O157:H7 dur-
ing the log-phase, as the pathogen took longer to reach the
maximum growth rate (M) in tomatoes (Fig. 2, Table 1). This
finding is in agreement with a study by Beuchat46 with ripe
tomatoes, which reported that the pH range of 3.9–4.4 present
in the ripe tomatoes prevents or retards bacterial growth. Del
Rosario and Beuchat47 also suggested that vegetables and fruit
juices with a pH value less than 4.0 are, in general, not good
substrates to support the growth of E. coli O157:H7.
Other researchers have shown that MPD decreases sig-
nificantly when the concentration of preservatives increases
or when natural microbial compounds are present.48 As in
the case of cucumber the volatile oils and the methanolic
and dichloromethane extracted from the peel of C. sativus
exhibited antimicrobial activity.49 Cho et al.,50 reported that
the high content of the volatiles such as 2,6-nonadienal and
2-nonenal show strong activity against E. coli O157:H7. In
contrast to this, there are no reports on the presence of
antimicrobial compounds against human pathogens in the
skin of tomato and onion. Although not specifically against
E. coli O157:H7, but still some studies have reported antimicro-
bial activity of onion aqueous extracts against gram-negative
microorganisms.51 However, Rounds et al.,52 showed that
powdered onion induced the growth of E. coli by 1.2 log CFU/g,
indicating that onion, like other vegetables, provides nutrients
that enhance the growth of bacteria. Nevertheless, there is no
conclusive explanation on the delay of the growth rate of E.
coli O157:H7 on those vegetables.
In conclusion, the predictive models presented in this
study offer a good comparative account of the growth of E. coli
O157:H7 on samples with different intrinsic factors demon-
strating low selectivity of the pathogen. The data suggest that
 b r a z i l i a n j o u r n a l o f m i c r o b i o l o g y 4 9 (2 0 1 8) 104–111 109

Table 1 – Mathematical modeling of the growth of E. coli O157:H7 adhered to the epicarp of vegetables and fruits. The
Gompertz (A, B, C, M) and derived parameters (GT, generation time; EGR, exponential growth rate; LPD, lag-phase
duration; MPD, maximum population density) are shown. Data are presented as mean ± standard error (n = 3). The letters
between the columns indicate significant differences (p < 0.05).
A (log CFU/mL) Bgrowth (h) C (log CFU/mL) M (h) GT (h) EGR (log LPD (h) MPD (log
CFU/mL/h) CFU/mL)

Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE Mean S.E Mean S.E

Tomato 2.819 0.12 a 1.498 0.60 a 2.060 0.59 a 3.631 0.03 a 0.398 0.11 a 0.881 0.23 a 2.895 1.53 a 4.279 0.54 a
Beet 2.139 0.12 b 1.684 1.30 a 2.729 1.20 a 2.009 0.38 b 0.562 0.03 a 0.540 0.03 a 2.230 1.50 a 4.868 0.27 b
Radish 2.507 0.15 a 1.431 0.63 a 2.537 0.80 a 2.032 0.13 b 0.336 0.07 a 0.968 0.18 a 1.026 0.37 a 5.044 0.69 b
Chili 2.573 0.15 a 1.344 0.92 a 2.351 1.00 a 2.066 0.55 b 0.632 0.07 a 0.491 0.06 a 2.709 1.30 a 4.924 1.05 b
Cucumber 2.742 0.05 a 1.790 0.84 a 1.971 0.79 a 3.351 0.17 a 0.413 0.10 a 0.806 0.16 a 1.966 0.69 a 4.314 0.85 a
Onion 2.804 0.03 a 1.286 0.59 a 1.921 0.61 a 2.961 0.06 a 0.480 0.06 a 0.647 0.08 a 2.193 0.74 a 4.125 0.63 a

F-value 5.193 0.053 0.147 3.766 1.953 1.847 0.349 4.052

p-value 0.009 0.998 0.977 0.045 0.159 0.178 0.873 0.048

Table 2 – Epicarp proximal composition (%) of vegetables Acknowledgement

and fruits (n = 5).
Moisture Ash Crude fiber pH The authors are grateful to Ana Ruth Christopher Ramos
from CINVESTAV-Mérida (México) for technical assistance
Mean SE Mean SE Mean SE Mean SE
with the scanning electron microscopy. We also thank Daniela
Tomato 95.4 0.23 0.74 0.04 0.57 0.22 4.77 0.08 A. Acevedo Ojeda from UMM for assistance with PCR analysis.
Beet 81.6 0.25 1.69 0.13 1.11 0.24 6.44 0.07
This project was supported by FOMIX-CONACYT – Gobierno
Radish 90.0 0.76 1.34 0.37 0.65 0.08 7.03 0.04
del Estado de Yucatán (México), grant no. 108103.
Chili 92.3 0.35 0.76 0.01 2.98 0.19 6.30 0.40
Cucumber 94.9 0.17 0.64 0.03 1.25 0.07 7.08 0.11
Onion 95.1 0.27 0.27 0.03 0.47 0.04 5.40 0.60

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depends on the intrinsic factors of the vegetables and fruits
under study, with the clear exception of tomato, cucumber and
onion, wherein the delay in the growth of the pathogen was
significant. However, it is important to note that the growth
models constructed under controlled environmental condi-
tions offer limited insights and may vary widely from the
observed profile of vegetables and fruits in the field, since
this study was designed to inoculate the samples with E. coli
O157:H7 without the presence of competitive microflora on
the surface. A lower concentration of the microorganism could
also be expected in the field than that was employed in the
study in the laboratory. The results reported by Brackett,52
suggested that reducing the native microbial populations by
washing and sanitizing or by controlled atmosphere storage,
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vegetables. The reduction in the surface populations reduces
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consumption. Future research in this field should aim to ver-
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O157:H7 in the tissues of vegetables under realistic field
conditions.
Conflicts of interest
The authors declare no conflicts of interest.
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